Proteomic Analysis Identifies Three Reliable Biomarkers of Intestinal Inflammation in the Stools of Patients With Inflammatory Bowel Disease

• Published in: Journal of Crohn's and colitis 2023-01, Vol.17 (1), p.92-102
• Main Authors: Vitali, Roberta, Palone, Francesca, Armuzzi, Alessandro, Fulci, Valerio, Negroni, Anna, Carissimi, Claudia, Cucchiara, Salvatore, Stronati, Laura
• Format: Article
• Language: English
• Subjects: Biomarkers - analysis; Chymotrypsin - metabolism; Colitis, Ulcerative - diagnosis; Colitis, Ulcerative - metabolism; Crohn Disease - diagnosis; Crohn Disease - metabolism; Feces - chemistry; Gelsolin - metabolism; Humans; Inflammation - metabolism; Inflammatory Bowel Diseases - complications; Inflammatory Bowel Diseases - diagnosis; Inflammatory Bowel Diseases - metabolism; Intestinal Mucosa - metabolism; Leukocyte L1 Antigen Complex - analysis; Proteomics; rho Guanine Nucleotide Dissociation Inhibitor beta - metabolism; Severity of Illness Index
• ISSN: 1873-9946; 1876-4479
• DOI: 10.1093/ecco-jcc/jjac110

Abstract Background Faecal biomarkers have emerged as important tools in managing of inflammatory bowel disease [IBD], which includes Crohn’s disease [CD] and ulcerative colitis [UC]. Aim To identify new biomarkers of gut inflammation in the stools of IBD patients using a proteomic approach. Methods Proteomic analysis of stools was performed in patients with both active CD and CD in remission and in controls by 2-DIGE and MALDI-TOF/TOF MS. An ELISA was used to confirm results in a second cohort of IBD patients and controls. Results 2-DIGE analysis detected 70 spots in the stools of patients with active CD or patients in remission CD and in controls. MALDI-TOF/TOF MS analysis identified 21 proteins with Chymotrypsin C, Gelsolin and Rho GDP-dissociation inhibitor 2 [RhoGDI2] best correlating with the levels of intestinal inflammation. Results were confirmed in a second cohort of IBD patients and controls [57 CD, 60 UC, 31 controls]. The identified faecal markers significantly correlated with the severity of intestinal inflammation in IBD patients [SES-CD in CD, Mayo endoscopic subscore in UC] [CD; Chymotrypsin-C: r = 0.64, p < 0.001; Gelsolin: r = 0.82, p < 0.001; RhoGDI2: r = 0.64, p < 0.001; UC; Chymotrypsin-C: r = 0.76, p < 0.001; Gelsolin: r = 0.75, p < 0.001; RhoGDI2: r = 0.63, p < 0.001]. Moreover, ROC analysis showed that Gelsolin [p < 0.0002] and RhoGDI2 [p < 0.0001] in CD, and RhoGDI2 [p = 0.0004] in UC, have higher sensitivity and specificity than faecal calprotectin in discriminating between patients and controls. Conclusions We show for the first time that 2-DIGE is a reliable method to detect proteins in human stools. Three novel faecal biomarkers of gut inflammation have been identified that display good specificity and sensitivity for identifying IBD and significantly correlate with IBD severity.